The BASHY Platform Enables the Assembly of a Fluorescent Bortezomib-GV1001 Conjugate

In this study, we show that fluorescent boronic-acid derived salicylidenehydrazone complexes (BASHY) can function as fluorescent linkers for bioconjugates that were used to monitor the delivery of the proteasome inhibitor bortezomib (Btz) to HT-29 cancer cells. BASHY complexes were structurally optimized to improve the stability of the complex in buffered conditions (ammonium acetate, pH 7 up to t(1/2) = 40 h), photophysically characterized regarding their fluorescence properties and used in confocal microscopy colocalization studies that revealed their intracellular sequestration by lipid droplets. The accumulation in these hydrophobic organelles limited the hydrolysis of the complex and consequently the drug release, a problem that was circumvented by the conjugation of the BASHY-Btz complex with a cell-penetrating peptide GV1001-C. The conjugate exhibited an improved cytoplasmic availability as confirmed by confocal fluorescence microscopy studies and an improved potency against HT-29 cancer cells (IC50 = 100 nM) as compared to the nontargeted complex (IC50 = 450 nM).

Automated summary

This study demonstrates that fluorescent boronic-acid derived salicylidenehydrazone complexes can be used as fluorescent linkers to monitor the delivery of proteasome inhibitor to cancer cells. The complexes were optimized for stability and sequestered by lipid droplets within cells, limiting hydrolysis and drug release. Conjugation with a cell-penetrating peptide improved cytoplasmic availability and potency against cancer cells.